LOCUS       sequence-247704-        3078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly AE destination vector DVCK-AE-GFP 
            (ColE1/kan resistance destination vector with constitutive GFP 
            expression for screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3078)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 3078)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: V35m
FEATURES             Location/Qualifiers
     source          1..3078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     497..517
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      576..647
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      663..690
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    800..820
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      821..878
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(902..919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1127..1146)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1138..1953
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1211..1230)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2075..2663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2155..2174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2704..2722
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2726..2769)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2772..2793
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2857..2891
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     CDS             join(2922..3078,1..557)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2952..2980)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tacctgttcc atggccaaca cttgtcacta ctttcggtta tggtgttcaa tgctttgcga
       61 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg
      121 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
      181 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
      241 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca
      301 tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag
      361 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
      421 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
      481 aaaagagaga tcacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
      541 tggatgaact atacaaataa taaaggtcca ggcatcaaat aaaacgaaag gctcagtcga
      601 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
      661 ctggctcacc ttcgggtggg cctttctgcg tttatagctt atgtcgttgc gctcactgcc
      721 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      781 ggtctctgct tatgtcttct actagtagcg gccgctgcag tccggcaaaa aagggcaagg
      841 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt
      901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      961 caaaggcggt aagtcgcgtg ttatacgccc gttatccatg ggtatggaca gttttccctt
     1021 tgatatgtaa cgcacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1081 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1141 agccatattc aacgggaaac gtcttgctcc cgtccgcgct taaactccaa catggacgct
     1201 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1261 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1321 gccaatgatg ttacagatga gatggtccgt ctcaactggc tgacggagtt tatgcctctc
     1381 ccgaccatca agcattttat ccgtactcct gatgatgcgt ggttactcac caccgcgatt
     1441 cctgggaaaa cagccttcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1501 gatgcgctgg ccgtgttcct gcgccggtta cattcgattc ctgtttgtaa ttgtcctttt
     1561 aacagcgatc gtgtatttcg tcttgctcag gcgcaatcac gcatgaataa cggtttggtt
     1621 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1681 atgcacaagc tcttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1741 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgggtcgga
     1801 atcgcagacc gttaccagga ccttgccatt ctttggaact gcctcggtga gttttctcct
     1861 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1921 cagtttcatt tgatgctcga tgagtttttc taataatact agcagaaatc atccttagcg
     1981 aaagctaagg atttttttta tctgattacc gcctttgagt gagcgtcgac ctagtcctag
     2041 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2101 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2161 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2221 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2281 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2341 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2401 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2461 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2521 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2581 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2641 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgcgc
     2701 ataaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     2761 gatgatttct ggaattcgcg gccgcttcta gagactagtg gaagacatgg agagagacct
     2821 gcaccatatg cggtgagact agtggaagac atggagttga cagctagctc agtcctaggt
     2881 actgtgctag ctactagagt cacacaggaa agtactaaat gatgcgtaaa ggagaagaac
     2941 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     3001 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
     3061 tttgcactac tggaaaac
//
